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枯草芽孢杆菌染色体的正常复制终点terC,对于营养生长和孢子形成来说并非必需。

The normal replication terminus of the Bacillus subtilis chromosome, terC, is dispensable for vegetative growth and sporulation.

作者信息

Iismaa T P, Wake R G

机构信息

Department of Biochemistry, University of Sydney, N.S.W., Australia.

出版信息

J Mol Biol. 1987 May 20;195(2):299-310. doi: 10.1016/0022-2836(87)90651-6.

Abstract

The Bacillus subtilis strains CU1693, CU1694 and CU1695 were shown by hybridization analysis to carry large deletions of the terminus region that originated within discrete fragments of the SP beta prophage genome. The absence of terC in CU1693 was demonstrated definitively by the identification of a novel junction fragment comprising SP beta DNA and DNA that lies on the other side of terC in the parent strain. This represented the deletion of approximately 230 kb of CU1693 DNA, with the removal of approximately 150 kb to the left of terC and approximately 80 kb to the right of terC. The lack of hybridization of CU1694 and CU1695 DNA to cloned DNA carrying the terC sequence and to cloned DNAs flanking terC suggested that terC is absent from the chromosome of each of these strains also, and that the deletions in CU1694 and CU1695 extend beyond the segment of the terminus region that has been mapped and cloned. The normal growth rate and morphology of CU1693, CU1694 and CU1695 relative to the parent strain when grown in complex medium indicated dispensability of terC for vegetative growth and division. B. subtilis SU153 was constructed using a specific deletion-insertion vector that was designed to effect the deletion of 11.2kb of DNA spanning terC, with the removal of approximately 9.7kb to the left of terC and approximately 1.kb to the right of terC. This manipulation did not introduce any readily detectable auxotrophic requirement. Physiological characterization of SU153 confirmed the dispensability of terC for vegetative growth and cell division, and also established the lack of requirement of terC for the specialized cell division that is associated with formation of the bacterial endospore.

摘要

通过杂交分析表明,枯草芽孢杆菌菌株CU1693、CU1694和CU1695携带末端区域的大片段缺失,这些缺失起源于SPβ原噬菌体基因组的离散片段内。通过鉴定一个包含SPβ DNA和亲本菌株中位于terC另一侧的DNA的新型连接片段,明确证实了CU1693中terC的缺失。这代表了约230 kb的CU1693 DNA缺失,terC左侧约150 kb和terC右侧约80 kb被去除。CU1694和CU1695 DNA与携带terC序列的克隆DNA以及terC侧翼的克隆DNA缺乏杂交,这表明这些菌株的染色体中也不存在terC,并且CU1694和CU1695中的缺失延伸到了已定位和克隆的末端区域片段之外。当在复杂培养基中生长时,CU1693、CU1694和CU1695相对于亲本菌株的正常生长速率和形态表明terC对于营养生长和分裂是可有可无的。枯草芽孢杆菌SU153是使用一种特定的缺失插入载体构建的,该载体设计用于实现跨越terC的11.2 kb DNA的缺失,terC左侧约9.7 kb和terC右侧约1 kb被去除。这种操作没有引入任何易于检测到的营养缺陷型需求。SU153的生理特性证实了terC对于营养生长和细胞分裂是可有可无的,并且还确定了terC对于与细菌芽孢形成相关的特殊细胞分裂没有需求。

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